Antioxidant effect of angiotensin (1-7) in the protection of pancreatic ß cell function

Zhang, Fen; Liu, Chang; Wang, Lei; Cao, Xi; Wang, Ying Ying; Yang, Jin Kui

MMP. MMP was assessed using JC-1 (Beyotime Institute of Biotechnology) in INS-1 cells. Cells grown in 6-well plates were incubated with 1 ml JC-1 (2 mg/ml) for 20 min at 37˚C. The cells were centrifuged at 1,000 x g for 5 min, while the pellets were resuspended in 500 µl PBS. Carbonyl cyanide M-chlorophenylhydrazone (CCCP; Beyotime Institute of Biotechnology), a mitochondrial electron transport chain inhibitor, served as a positive control. CCCP (10 M) was added to the cells for 20 min. In total, 10,000 cells from each well were analyzed by flow cytometry (ex, 490 nm, em, 590 nm). Color change from green to red indicated an increase in the MMP or the improvement of mitochondrial function.